IMPROVED METHODS FOR IMMUNO-ELECTRON MICROSCOPY OF CULTURED CELLS: USE OF A NOVEL SUBSTRATE AND APPLICATION OF MICROWAVE IRRADIATION

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For immuno-electron-microscopic observations of cultured cells, we used a novel substrate, a commonly used domestic wrapping film, for plating cells, and irradiated microwave (MW) during fixation or during post-embedding immunoreaction. Isolated hepatocytes cultured on wrapping film could be observed under a phase-contrast microscope, appeared healthy, and were sectioned invarious orientations for electron microscopy. Post-embedding immunogold staining was carried out on these sections, and the density of gold particles was used to estimate the effects of MW irradiation. Although MY irradiation (60 sec) during fixation brought about a comparable density to that of conventional fixation (15-min immersion), the preservation of fine structures was poorer in irradiated specimens. The time required for the reaction with primary antibody was reduced with MW irradiation. Sixty-second irradiation was shown to give an equivalent result to overnight reaction without irradiation.
日本組織細胞化学会の論文